Barcoding Neotropical Psittacidae

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Barcoding Neotropical Psittacidae
ABBI Regional Working Group Meeting Buenos Aires,
14-16 March 2007 Cristina Y. Miyaki Departamento
de Genética e Biologia Evolutiva Instituto de
Biociências Universidade de São Paulo
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Psittaciformes distribution
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Monophyly of Arini Bayesian unrooted (3062 bp
12S, 16S intron 5 tropomyosin) Red
Neotropical Blue African Green
Australasian Bold no MP support, only Bayesian
Matsumoto et al, prep.
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Photo P.J. Faria
Arini 150 species 27 endangered
Source Forshaw 1989
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Species identification of non hatched embryos
Application of DNA barcodes in Neotropical
Psittacidae
Which species?
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Species identification of non hatched embryos
Choice of marker mitochondrial 16S rDNA
Sequenced 141-509 bp 6 groups
(haplotypes) Analyses BLAST (GenBank) and
distance
Matsumoto, Oliveira-Marques, et al., in prep.
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213 bp of 16S rDNA Ara ararauna (3)
Non psitacid (1)
Matsumoto, Oliveira-Marques, et al., in prep.
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390 bp of 16S rDNA Ara ararauna (3)
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Species identification of non hatched
embryosConclusions

• The available 16S rDNA data base was informative
  to identify the species of the embryos.
• The species identification was confirmed in
  another lab that used another mitochondrial
  marker (cytochrome b).

Would COI be a good marker? Building a COI
database for Neotropical parrots and will test
the same embryos.
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COI sequence database for Neotropical parrots

• Masters Degree student Priscila F.M. Gonçalves
  (Dec 2006)
• Objective construct the database and evaluate
  its usefulness for species identification (e.g.
  evaluate if genetic distances within and among
  taxa do not overlap)

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COI sequence database for Neotropical parrots

• Samples
• 113 out of 150 species of Neotropical parrots (34
  with skins)

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COI sequence database for Neotropical parrots

• PCR conditions (touchdown)
• 5 min 95ºC
• 14 cycles 30 sec 95ºC, 30 sec 68ºC (1C lower
  per cycle), 40 sec 72ºC
• 29 cycles 30 sec 95, 30 sec 54C, 40 sec 72C
• 10 min 72C
• Sequences obtained 23 (March 2)
• Average size of sequences 792 bp (732-853 bp)

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Acknowledgements
Erika Tavares, Royal Ontario Museum, Toronto